A combined phage display ScFv library against Myxobolus rotundus infecting crucian carp, Carassius auratus auratus (L.), in China

Immunological methods have been developed for the diagnosis of Myxobolus rotundus but their use has been limited for the prevention and therapy of this serious parasitic pathogen. Phage display antibody libraries are a powerful technique for the development of antibodies to molecules of interest and have advantages over traditional hybridroma approaches. In the present study, four antigen fractions related to M. rotundus were prepared and a combined phage display single-chain antibody fragments (ScFv) library was constructed against this parasite. Preliminary analysis indicated that a combined antibody library of about 2.08 X 10(5) individual clones and high diversity was generated. After four rounds of screening (bio-panning) against soluble spore protein prepared from lysed, intact, mature M rotundus spores, a strain monoclonal phage display ScFv, termed pCAN-6H9, with better affinity, was isolated. The pCAN-6H9 gene fragment was sequenced and analysed. The specificity of pCAN-6H9 was further demonstrated by dot-blot. In competition enzyme-linked immunosorbent assay, both the original and enriched phage-displayed ScFv repertoire showed significant inhibition of mouse anti-M rotundus serum binding to coated antigen, while the inhibition rate of monoclonal pCAN-6H9 phage particles was only 11.83%.

Detection of Myxobolus rotundus (Myxozoa : Myxosporea) in skin mucus of crucian carp Carassius auratus auratus using a monoclonal antibody

Diagnosis of myxosporean Myxobolus rotundus infection was conducted by examining skin mucus from the infected crucian carp Carassius auratus auratus with a monoclonal antibody, MAb 2D12, raised previously against the parasite. A positive reaction was observed in skin mucus collected from infected fish, and spores and pre-spore stages of the parasite were identified by the MAb 2D12. It was also demonstrated that M. rotundus infection can be successfully detected by a simple method, enzyme-linked immunosorbent assay (ELISA), and that skin mucus collected from infected fish skin had a significantly higher optical density (OD) value than that from uninfected fish.

Efficiency of constructed wetlands in decontamination of water polluted by heavy metals

A twin-shaped constructed wetland (CW) comprising a vertical flow (inflow) chamber with Cyperus alternifolius followed by a reverse-vertical flow (outflow) chamber with Villarsia exaltata was assessed for decontamination of artificial wastewater polluted by heavy metals. After application of Cd, Cu, Pb, Zn over 150 days, together with Al and Mn during the final 114 days, no heavy metals with the exception of Mn could be detected in either the drainage zone at the bottom, shared by both chambers, or in the effluent. The inflow chamber was, therefore, seen to be predominantly responsible for the decontamination process of more toxic metal species with final concentrations far below WHO drinking-water standards. About one-third of the applied Cu and Mn was absorbed, predominantly by lateral roots of C. alternifolius. Lower accumulation levels were observed for Zn (5%), Cd (6%), Al (13%). and Pb (14%). Contents of Cd, Cu, Mn, and Zn in soil were highest in top layer, while Al and Pb were evenly distributed through the whole soil column. Metal species accumulating mainly in the top layer can be removed mechanically. A vertical flow CW with C. alternifolius is an effective tool in phytoremediation for treatment of water polluted with heavy metals. (C) 2002 Elsevier Science B.V. All rights reserved.

西南极南设得兰群岛中新生代火山岩基底沉积学及其大地构造制约

Livingston Island, the second island of South Shetland Island, constains Mesozoic-Cenozoic basement, Mesozoic-Cenozoic volcanic sequences, plutonic intrusions and post-subduction volcanic rocks, which document the history and evolution of an important part of the South Shetland Islands magmatic arc. The sedimentary sequence is named the Miers Bluff Formation (MBF) and is interpreted as turbidite since the first geological study on South Shetland Islands, and is interpreted as turbidite. It base and top are not exposed, but a thickness of more than 3000m has been suggested and seems plausible. The turbidite is overlain by Mid - Cretaceous volcanic rocks and intruded by Eocene tonalites. The age of the Miers Bluff Formation is poorly constrained Late Carboniferous -Early Triassic. Sedimentary Environment, tectonic setting and forming age of sedimentary rocks of the Miers Bluff Formation were discussed by means of the methods of sedimentology, petrography and geochemistry, combinig with the study of trace fossils and microfossil plants. The following conclusions are obstained. A sedimentary geological section of Johnsons Dock is made by outside measuring and watching, and then according the section, the geological map near the Spanish Antarctic station was mapped. Four pebbly mudstone layers are first distinguished, which thickness is about 10m. The pebbly mudstone is the typical rock of debris flow, and the depostional environment of pebbly mudstone may be the channel of mid fan of submarine fan. The sedimentsry structural characteristics and size analysis of sandstones show the typical sedimentary feature of turbidity flow and the Miers Bluff Formation is a deep-water turbidite (include some gravity-flow sediments). The materials of palaeocurrents suggest the continental slope dip to southeast, and indicate the provenance of turbidity sediment in the northwest area. By facies analysis, six main facies which include seven subfacies were recognized, which are formed in mid-fan and lower-fan of submarine, meanwhile, the sedimentary features of each facies and subfacies are summarized. The study of clastic composition, major elements, trace elements and rare earth elements indicates the forming setting of the Miers Bluff Formaton is active continental margin and continental island arc and the provenance is dissected magmatic arc which main composition is felsic gneiss. Many trace fossils of the whole succession were found in the turbidites of the Miers Bluff Formation. All these trace fossils are deep sea ichnofossils. There are fifteen ichnogenus, sixteen ichnospecies. Moreover, a new trace fossil was found and a new ichnogenus and new ichnospecies was proposed - Paleaichnus antarctics ichnogen, et ichnosp, nov.. Except the new ichnogenus and ichnospecies, others had been found in deep-sea flysch turbidites. Some are in mudstone and are preserved in the cast convex of overlying sandstone sole, they formed before turbidity flows occurred and belong to the high-different Graphoglyptida of fiysch mudstone. Others as Fucusopsis and Neonereites are preserved in sandstones and stand for trace assemblages after turbidity sedimentation. These trace fossils are typical members of abyssal "Nereites" ichnofacies, and provide for the depositional environment of the Miers Bluff Formation. Fairly diverse microfossil plants have been recovered from the Miers Bluff Formation, Livingston Island, including spores, pollen, acritarchs, wood fragments and cuticles. Containing a total of about 45 species (forms) of miospores, the palynofiora is quantitatively characterized by the dominance of non-striate bisaccate pollen, but spores of pteridophytes and pollen of gymnosperms are proportionate in diversity. It is somewhat comparable to the subzone C+D of the Alisporites zone of Antarctica, and the upper Craterisporites rotundus zone and the lower Polycingulatisporites crenulatus zone of Australia, suggesting a Late Triassic (possibly Norian-Rhaetian) age, as also evidenced by the sporadic occurrence of Aratrisporites and probable Classopollis as well as the complete absence of bisaccate Striatiti. The parent vegetation and paleoclimate are preliminarily deduced. At last, the paper prooses the provenance of sedimentary rocks of the Miers Bluff Formation locates in the east part to the southern Chile(or Southern South American). In the Triassic period, contrasting with New Zealand, Australia and South American of the Pacific margin of Gondwanaland, the Miers Bluff Formation is deposited in the fore-arc basin or back-arc basin of magmatic arc.